稻瘟病菌66 kDa糖蛋白激发子的纯化及性质研究

 采用离心、超滤、柱层析等方法对稻瘟病菌(Magnaporthe grisca) ZC₁₃菌株97-151a细胞壁来源的水溶性糖蛋白激发子进行了纯化;经SDS-PAGE检测,GP66达到电泳纯;凝胶过滤法及SDS-PAGE测定表明,GP66分子量为64~66 kDa;经蒽酮-硫酸法检测,糖/蛋白比例为3.84;可诱导水稻叶片过氧化物酶与苯丙氨酸解氨酶活性升高。经热、胰蛋白酶和过碘酸钠处理后的生物活性检测结果表明,GP66的活性位点为糖基部分。

Purification and characterization of an elicitor (GP66) from Magnaporthe grisea

A water soluble elicitor (GP66) was isolated and purified from hyphal cell wall of the strain 97-151a of Magnaporthe grisea race ZC₁₃ through centrifugation, ultra-filtration and chromatography. SDS-PAGE showed that the elicitor GP66 had only one band of 66 kDa. Sephadex G-100 gel filtration showed that molecular weight of the elicitor was 64 kDa. Anthrone-colorimetric assay and Coomassie blue G-250 staining showed that the carbohydrate and protein contents were in a ratio of 3.84. Bioassay detection indicated that GP66 could increase the levels of the activity of POD and PAL in rice leaves. Digestion with trypsin and boiling at 100℃ for 10 min did not abolish the elicitor activity, but complete loss of activity was observed after periodate treatment. The results suggest that the carbohydrate moiety contains the active site of the elicitor, and the protein moiety may not be necessary for elicitor activity.